Identification and characterisation of Gamma-herpesviruses in zoo artiodactyla.

BACKGROUND: Viruses within the γ-herpesviruses subfamily include the causative agents of Malignant Catarrhal Fever (MCF) in several species of the order Artiodactyla. MCF is a usually fatal lymphoproliferative disease affecting non-adapted host species. In adapted host species these viruses become latent and recrudesce and transmit during times of stress or immunosuppression. The undetected presence of MCF-causing viruses (MCFVs) is a risk to non-adapted hosts, especially within non-sympatric zoological collections. This study investigated the presence of MCFVs in six different zoological collections in the UK, to evaluate the presence of subclinical/latent MCFVs in carrier animals. METHODS: One-hundred and thirty eight samples belonging to 54 different species of Artiodactyla were tested by Consensus Pan-herpes PCR. The positive samples were sequenced and subjected to phylogenetic analyses to understand their own evolutionary relationships and those with their hosts. RESULTS: Twenty-five samples from 18 different species tested positive. All viruses but one clustered in the γ-herpesvirus family and within the Macavirus as well as the non-Macavirus groups (caprinae and alcelaphinae/hippotraginae clusters, respectively). A strong association between virus and host species was evident in the Macavirus group and clustering within the caprinae group indicated potential pathogenicity. CONCLUSION: This study shows the presence of pathogenic and non-pathogenic MCFVs, as well as other γ-herpesviruses, in Artiodactyla species of conservation importance and allowed the identification of new herpesviruses in some non-adapted species.

Cancer Prevalence Across Vertebrates.

Cancer is pervasive across multicellular species, but what explains differences in cancer prevalence across species? Using 16,049 necropsy records for 292 species spanning three clades (amphibians, sauropsids and mammals) we found that neoplasia and malignancy prevalence increases with adult weight (contrary to Peto's Paradox) and somatic mutation rate, but decreases with gestation time. Evolution of cancer susceptibility appears to have undergone sudden shifts followed by stabilizing selection. Outliers for neoplasia prevalence include the common porpoise (<1.3%), the Rodrigues fruit bat (<1.6%) the black-footed penguin (<0.4%), ferrets (63%) and opossums (35%). Discovering why some species have particularly high or low levels of cancer may lead to a better understanding of cancer syndromes and novel strategies for the management and prevention of cancer.

Cancer Prevalence Across Vertebrates.

Cancer is pervasive across multicellular species. Are there any patterns that can explain differences in cancer prevalence across species? Using 16,049 necropsy records for 292 species spanning three clades (amphibians, sauropsids and mammals) we found that neoplasia and malignancy prevalence increases with adult weight and decreases with gestation time, contrary to Peto’s Paradox. Evolution of cancer susceptibility appears to have undergone sudden shifts followed by stabilizing selection. Outliers for neoplasia prevalence include the common porpoise (<1.3%), the Rodrigues fruit bat (<1.6%) the black-footed penguin (<0.4%), ferrets (63%) and opossums (35%). Discovering why some species have particularly high or low levels of cancer may lead to a better understanding of cancer syndromes and novel strategies for the management and prevention of cancer.

Somatic mutation rates scale with lifespan across mammals.

The rates and patterns of somatic mutation in normal tissues are largely unknown outside of humans1-7. Comparative analyses can shed light on the diversity of mutagenesis across species, and on long-standing hypotheses about the evolution of somatic mutation rates and their role in cancer and ageing. Here we performed whole-genome sequencing of 208 intestinal crypts from 56 individuals to study the landscape of somatic mutation across 16 mammalian species. We found that somatic mutagenesis was dominated by seemingly endogenous mutational processes in all species, including 5-methylcytosine deamination and oxidative damage. With some differences, mutational signatures in other species resembled those described in humans8, although the relative contribution of each signature varied across species. Notably, the somatic mutation rate per year varied greatly across species and exhibited a strong inverse relationship with species lifespan, with no other life-history trait studied showing a comparable association. Despite widely different life histories among the species we examined-including variation of around 30-fold in lifespan and around 40,000-fold in body mass-the somatic mutation burden at the end of lifespan varied only by a factor of around 3. These data unveil common mutational processes across mammals, and suggest that somatic mutation rates are evolutionarily constrained and may be a contributing factor in ageing.

Diagnosis and Treatment of Heavy Metal Toxicosis in Six Waldrapp Ibis (Geronticus eremita).

Six adult Waldrapp ibis (Geronticus eremita) were presented with heavy metal toxicosis secondary to paint chip ingestion after being placed in an indoor confinement enclosure because of a regional highly pathogenic avian influenza epornitic and government restrictions. Four of the ibis developed clinical disease signs between 45 and 64 days of confinement, including weakness, lethargy, inability to fly, low body condition, and bilateral wing droop while 2 birds appeared to remain normal during that period. Common clinicopathological findings included elevated plasma aspartate aminotransferase, creatine kinase, glutamate dehydrogenase, and bile acids values. Erythrocyte morphological changes included prominent hypochromasia, decreased size of polychromatophils with occasional cytoplasmic stippling, abnormal shapes, and D cells. Whole-body radiographic imaging revealed particulate radio-opaque material in the ventriculus of all affected birds. One bird died before chelation therapy could be instituted and heavy metal testing of the liver revealed increased concentrations of lead and zinc. Lead toxicosis was diagnosed in the 5 remaining birds by plasma lead analysis. These 5 birds were hospitalized for an average of 40 days and treated with sodium calcium edetate and fluid therapy, which resulted in resolution of the toxicosis. Plasma biochemistry, complete blood counts, and blood lead values were performed repeatedly on all 5 birds and were used to guide therapy and monitor treatment response. This case series emphasizes the importance of early contingency planning and reviews the treatment of lead toxicosis in birds with a focus on long-term clinical monitoring and hematology.

FERLAVIRUS-RELATED DEATHS IN A COLLECTION OF VIPERID SNAKES.

Between June and October 2013, 26 snakes of six viperid species kept in two adjoining rooms died ( n = 16) or were euthanized on medical (1) or welfare grounds (9). Two were from the main zoo collection, but the other 24 had been imported and quarantined for a minimum of 6 mo. Four of those that died and the single snake euthanized on medical grounds showed minor signs of respiratory disease prior to death, and five were weak, lethargic, and/or poor feeders. Frequent postmortem findings among all snakes were poor body condition (18) and respiratory disease (13). Seventeen cases were examined histologically, and pneumonia, sometimes with air sacculitis and/or tracheitis, was present in 15 individuals. Lung samples from 24 snakes were ferlavirus polymerase chain reaction (PCR) positive, and one of the two snakes for which only liver was available was also positive. The negative liver sample was from a snake that died of sepsis following anesthesia for surgical removal of a spindle cell sarcoma. Correlation with antemortem PCR testing of glottal and cloacal swabs in five cases was poor (sensitivity = 40%). Immunohistochemistry (IHC) for ferlaviruses on the tissues of 13 PCR-positive cases showed positive labeling in 7 only. Tissues samples from 22 ferlavirus PCR-positive snakes were examined for Chlamydia species by PCR, and 9 were positive, although DNA sequencing only confirmed two of three tested as Chlamydia pneumoniae. Immunohistochemistry for Chlamydia pneumoniae of seven cases (two Chlamydiales PCR positive, one of which was sequenced as C. pneumoniae, plus five negative) confirmed the Chlamydia PCR results. These two Chlamydiales PCR and IHC positive snakes were ferlavirus PCR positive, but IHC negative suggesting that, even though a ferlavirus was the predominant cause of the outbreak, in a few cases death may have been due to chlamydiosis with ferlavirus present, but not acting as the primary pathogen.

OSTEOCHONDROSIS IN THE DISTAL FEMURS OF AN ADULT RETICULATED GIRAFFE (GIRAFFA CAMELOPARDALIS RETICULATA): MACROSCOPIC, RADIOLOGIC, AND HISTOLOGIC FINDINGS.

An adult male reticulated giraffe (Giraffa camelopardalis reticulata) was presented for postmortem examination. During radiologic examination of the hindlimbs, osseous cyst-like lesions were detected in both medial femoral condyles. These lesions were subsequently examined macroscopically and histologically. The gross appearance suggested a diagnosis of bilateral osteochondrosis that was confirmed with histopathologic examination. This finding has not previously been reported in giraffes. Macroscopic visualization of the major limb joints, including the femorotibial joints, is therefore encouraged in future postmortem examinations of giraffes (Giraffa camelopardalis), and further assessment of clinical significance is required.

A novel investigation of a blister-like syndrome in aquarium Echinopora lamellosa.

This study investigates potential causes of a novel blister-like syndrome in the plating coral Echinopora lamellosa. Visual inspections of this novel coral syndrome showed no obvious signs of macroparasites and the blisters themselves manifested as fluid-filled sacs on the surface of the coral, which rose from the coenosarc between the coral polyps. Histological analysis of the blisters showed that there was no associated necrosis with the epidermal or gastrodermal tissues. The only difference between blistered areas and apparently healthy tissues was the presence of proliferated growth (possible mucosal cell hyperplasia) directly at the blister interface (area between where the edge of the blister joined apparently healthy tissue). No bacterial aggregates were identified in any histological samples, nor any sign of tissue necrosis identified. We conclude, that the blister formations are not apparently caused by a specific microbial infection, but instead may be the result of irritation following growth anomalies of the epidermis. However, future work should be conducted to search for other potential casual agents, including viruses.

Pituitary pars intermedia dysfunction (Equine Cushing's disease) in an onager (Equus hemionus onager).

Pituitary pars intermedia dysfunction (PPID), also known as Equine Cushing's disease, is most often diagnosed in older horses and ponies. To the authors' knowledge, there have been no reports of its diagnosis in captive nondomestic equids. A 13-yr old onager (Equus hemionus onager) at the Zoological Society of London (ZSL) Whipsnade Zoo showed clinical signs suggestive of PPID, including hirsutism, fat redistribution, weight loss, laminitis, and chronic infections. A dexamethasone suppression test was performed to confirm PPID. Subsequently, adenomatous hyperplasia and microadenoma of the pars intermedia were identified postmortem. Four months later, this onager's dam died suddenly, and adenomatous hyperplasia of the pars intermedia was diagnosed following necropsy. The dam had shown no clinical signs of PPID. Examination of archives identified eight other adult onagers that died or were euthanized between 1993 and 2007. The brain was not examined in four of these, but pituitary glands were described as enlarged during necropsy in three animals based on the subjective assessment of an experienced zoo and wildlife pathologist, making an overall prevalence of enlargement of 83.3%. Hyperplastic pituitary changes are positively correlated with age in domestic equids, and this may also be the case in onagers. Alternative etiologies are also discussed.

Analysis of biochemical markers of bone metabolism in Asian elephants (Elephas maximus).

Two human enzyme immunoassays (EIA) and one radioimmunoassay (RIA) were validated and used to measure osteocalcin (OC), bone alkaline phosphatase (BAP), and the cross-linked telopeptide domain of type I collagen (ICTP), in serum from Asian elephants (Elephas maximus). Sera from four adult females sampled on 7 consecutive days were also analyzed to assess the existence and magnitude of intraindividual day-to-day variability of the serum concentration of these markers. Sample dilution curves were parallel with assay standard curves, which demonstrated that excellent cross reactivity existed between assay antibodies and elephants marker antigens. Statistically significant inverse correlations were found between age and concentrations of all three markers: BAP, r = -0.862 (P < 0.01); OC, r = -0.788 (P < 0.002); and ICTP, r = -0.848 (P < 0.01). Strong positive correlations were found between BAP and OC (r = 0.797, P < 0.01), OC and ICTP (r = 0.860, P < 0.01), and between BAP and ICTP (r = 0.958, P < 0.01). No statistically significant intraindividual variability was found over 7 days in the four adult females for any of the markers assessed (OC: P = 0.089; ICTP: P = 0.642; BAP: P = 0.146; n=4 in each case). The overall coefficient of variability observed in this group of animals was 10.3%, 7.4%, and 5.5% for OC, BAP, and ICTP, respectively. These results suggest a potential role for biochemical markers of bone turnover in monitoring skeletal health and bone disease in Asian elephants.

A novel subgroup of rhadinoviruses in ruminants.

In the course of investigating the malignant catarrhal fever (MCF) subgroup of rhadinoviruses, seven novel rhadinoviruses were identified in a variety of ruminants, including domestic sheep, bighorn sheep, bison, black-tailed deer, mule deer, fallow deer, elk and addax. Based on the DNA polymerase gene sequences, these newly recognized viruses clustered into a second distinct subgroup in ruminants with three members identified previously in cattle, goats and oryx. Phylogenetic analysis revealed that the currently known ruminant rhadinoviruses appear to comprise three distinct genetic lineages: (i) the MCF subgroup, defined by sequence identity and the presence of the 15A antigenic epitope; (ii) a second distinct subgroup, devoid of the 15A epitope, which contains the previously reported bovine lymphotropic herpesvirus and related viruses; and (iii) a third distinct subgroup represented by Bovine herpesvirus 4. Comparison of phylogenetic trees between the rhadinoviruses and their corresponding hosts further supports the gammaherpesvirus and host co-evolution theory.

Symblepharon with aberrant protrusion of the nictitating membrane in the snowy owl (Nyctea scandiaca).

Two young snowy owl chicks were presented with aberrant protrusion of the nictitating membranes. This was caused by conjunctival adhesions causing symblepharon secondary to a previous septicemia episode. While symblepharon has been noted in birds before, this unusual presentation of the nictitating membrane has not been reported. Surgical intervention ameliorated the clinical signs, allowing vision in one bird by removal of the nictitating membranes, a technique which appeared to have no deleterious effects on the ocular surface.